LOCUS       sequence-432848-        9918 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Variant CE1 construct with BDFV HUH endonuclease, expressed from CMV
            or T7 promoters..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9918)
  AUTHORS   Ferreira da Silva J, Tou CJ, King EM, Eller ML, Rufino-Ramos D, Ma 
            L, Cromwell CR, Metovic J, Benning FMC, Chao LH, Eichler FS, 
            Kleinstiver BP
  TITLE     Click editing enables programmable genome writing using DNA 
            polymerases and HUH endonucleases.
  JOURNAL   Nat Biotechnol. 2024 Jul 22. doi: 10.1038/s41587-024-02324-x.
  PUBMED    39039307
REFERENCE   2  (bases 1 to 9918)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Biotechnol. 2024 Jul 22. doi: 10.1038/s41587-024-02324-x."
COMMENT     Sequence Label: HUHe variant click editor (CE1) -
            pCMV-T7-BDFV...
FEATURES             Location/Qualifiers
     source          1..9918
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(113..217)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(296..315)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        487..866
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        867..1070
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1021..1041
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1112..1131
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1112..1130
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(1576..4089,4090..4092,4093..5676)
                     /codon_start=1
                     /product="nickase mutant of the Cas9 endonuclease from the 
                     Streptococcus pyogenes Type II CRISPR/Cas system"
                     /label=Cas9(H840A)
                     /note="generates RNA-guided single strand nicks in DNA due 
                     to the H840A mutation in the HNH catalytic domain"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             1576..5676
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             5734..5754
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7639..7659
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7668..7685
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(7708..7725)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7714..7938
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(8009..8025)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8009..8025)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8022..8044)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8033..8049
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(8057..8063,8064..8081,8082..8087))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8102..8123
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8240..8257)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8411..8999)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8491..8510)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9170..9918,1..43,44..112))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     9793..9812
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
       61 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
      121 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
      181 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
      241 ctgacgtcga cggatcggga gatcgatctc ccgatcccct agggtcgact ctcagtacaa
      301 tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg
      361 ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca
      421 tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata
      481 cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc
      541 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
      601 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
      661 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
      721 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
      781 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
      841 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
      901 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
      961 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
     1021 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga
     1081 accgtcagat ccgctagaga tccgcggccg ctaatacgac tcactatagg gagagccgcc
     1141 accatgtccc cagaagatca cacccgcccc ggcacaatcc ccagcaagga aggctctggc
     1201 tgtagaagat ggtgcttcac cctgaacaat cctacagacg gcgaaatcga gttcgtgcgg
     1261 agcctgggac ctgacgagtt ctactacgcc atcgtcggca gagagaaagg cgagcagggc
     1321 acccctcacc tccagggata tttccacttt aagaacaaga agcggctgag cgccctgaag
     1381 aaaatgctgc ctagaggcca cttcgagcgg gccaagggca gcgacgccga caacgagaag
     1441 tactgcagca aggaaggaga tgtgatcctg accctgggca tcgtggctcg ggacggccat
     1501 agaagcggag gctctagcgg ctccgagaca cccggcacat ctgagtccgc cacccctgag
     1561 agcagcggcg gcagcgacaa gaagtacagc atcggcctgg acatcggcac caactctgtg
     1621 ggctgggccg tgatcaccga cgagtacaag gtgcccagca agaaattcaa ggtgctgggc
     1681 aacaccgacc ggcacagcat caagaagaac ctgatcggag ccctgctgtt cgacagcggc
     1741 gaaacagccg aggccacccg gctgaagaga accgccagaa gaagatacac cagacggaag
     1801 aaccggatct gctatctgca agagatcttc agcaacgaga tggccaaggt ggacgacagc
     1861 ttcttccaca gactggaaga gtccttcctg gtggaagagg ataagaagca cgagcggcac
     1921 cccatcttcg gcaacatcgt ggacgaggtg gcctaccacg agaagtaccc caccatctac
     1981 cacctgagaa agaaactggt ggacagcacc gacaaggccg acctgcggct gatctatctg
     2041 gccctggccc acatgatcaa gttccggggc cacttcctga tcgagggcga cctgaacccc
     2101 gacaacagcg acgtggacaa gctgttcatc cagctggtgc agacctacaa ccagctgttc
     2161 gaggaaaacc ccatcaacgc cagcggcgtg gacgccaagg ccatcctgtc tgccagactg
     2221 agcaagagca gacggctgga aaatctgatc gcccagctgc ccggcgagaa gaagaatggc
     2281 ctgttcggaa acctgattgc cctgagcctg ggcctgaccc ccaacttcaa gagcaacttc
     2341 gacctggccg aggatgccaa actgcagctg agcaaggaca cctacgacga cgacctggac
     2401 aacctgctgg cccagatcgg cgaccagtac gccgacctgt ttctggccgc caagaacctg
     2461 tccgacgcca tcctgctgag cgacatcctg agagtgaaca ccgagatcac caaggccccc
     2521 ctgagcgcct ctatgatcaa gagatacgac gagcaccacc aggacctgac cctgctgaaa
     2581 gctctcgtgc ggcagcagct gcctgagaag tacaaagaga ttttcttcga ccagagcaag
     2641 aacggctacg ccggctacat tgacggcgga gccagccagg aagagttcta caagttcatc
     2701 aagcccatcc tggaaaagat ggacggcacc gaggaactgc tcgtgaagct gaacagagag
     2761 gacctgctgc ggaagcagcg gaccttcgac aacggcagca tcccccacca gatccacctg
     2821 ggagagctgc acgccattct gcggcggcag gaagattttt acccattcct gaaggacaac
     2881 cgggaaaaga tcgagaagat cctgaccttc cgcatcccct actacgtggg ccctctggcc
     2941 aggggaaaca gcagattcgc ctggatgacc agaaagagcg aggaaaccat caccccctgg
     3001 aacttcgagg aagtggtgga caagggcgct tccgcccaga gcttcatcga gcggatgacc
     3061 aacttcgata agaacctgcc caacgagaag gtgctgccca agcacagcct gctgtacgag
     3121 tacttcaccg tgtataacga gctgaccaaa gtgaaatacg tgaccgaggg aatgagaaag
     3181 cccgccttcc tgagcggcga gcagaaaaag gccatcgtgg acctgctgtt caagaccaac
     3241 cggaaagtga ccgtgaagca gctgaaagag gactacttca agaaaatcga gtgcttcgac
     3301 tccgtggaaa tctccggcgt ggaagatcgg ttcaacgcct ccctgggcac ataccacgat
     3361 ctgctgaaaa ttatcaagga caaggacttc ctggacaatg aggaaaacga ggacattctg
     3421 gaagatatcg tgctgaccct gacactgttt gaggacagag agatgatcga ggaacggctg
     3481 aaaacctatg cccacctgtt cgacgacaaa gtgatgaagc agctgaagcg gcggagatac
     3541 accggctggg gcaggctgag ccggaagctg atcaacggca tccgggacaa gcagtccggc
     3601 aagacaatcc tggatttcct gaagtccgac ggcttcgcca acagaaactt catgcagctg
     3661 atccacgacg acagcctgac ctttaaagag gacatccaga aagcccaggt gtccggccag
     3721 ggcgatagcc tgcacgagca cattgccaat ctggccggca gccccgccat taagaagggc
     3781 atcctgcaga cagtgaaggt ggtggacgag ctcgtgaaag tgatgggccg gcacaagccc
     3841 gagaacatcg tgatcgaaat ggccagagag aaccagacca cccagaaggg acagaagaac
     3901 agccgcgaga gaatgaagcg gatcgaagag ggcatcaaag agctgggcag ccagatcctg
     3961 aaagaacacc ccgtggaaaa cacccagctg cagaacgaga agctgtacct gtactacctg
     4021 cagaatgggc gggatatgta cgtggaccag gaactggaca tcaaccggct gtccgactac
     4081 gatgtggacg ctatcgtgcc tcagagcttt ctgaaggacg actccatcga caacaaggtg
     4141 ctgaccagaa gcgacaagaa ccggggcaag agcgacaacg tgccctccga agaggtcgtg
     4201 aagaagatga agaactactg gcggcagctg ctgaacgcca agctgattac ccagagaaag
     4261 ttcgacaatc tgaccaaggc cgagagaggc ggcctgagcg aactggataa ggccggcttc
     4321 atcaagagac agctggtgga aacccggcag atcacaaagc acgtggcaca gatcctggac
     4381 tcccggatga acactaagta cgacgagaat gacaagctga tccgggaagt gaaagtgatc
     4441 accctgaagt ccaagctggt gtccgatttc cggaaggatt tccagtttta caaagtgcgc
     4501 gagatcaaca actaccacca cgcccacgac gcctacctga acgccgtcgt gggaaccgcc
     4561 ctgatcaaaa agtaccctaa gctggaaagc gagttcgtgt acggcgacta caaggtgtac
     4621 gacgtgcgga agatgatcgc caagagcgag caggaaatcg gcaaggctac cgccaagtac
     4681 ttcttctaca gcaacatcat gaactttttc aagaccgaga ttaccctggc caacggcgag
     4741 atccggaagc ggcctctgat cgagacaaac ggcgaaaccg gggagatcgt gtgggataag
     4801 ggccgggatt ttgccaccgt gcggaaagtg ctgagcatgc cccaagtgaa tatcgtgaaa
     4861 aagaccgagg tgcagacagg cggcttcagc aaagagtcta tcctgcccaa gaggaacagc
     4921 gataagctga tcgccagaaa gaaggactgg gaccctaaga agtacggcgg cttcgacagc
     4981 cccaccgtgg cctattctgt gctggtggtg gccaaagtgg aaaagggcaa gtccaagaaa
     5041 ctgaagagtg tgaaagagct gctggggatc accatcatgg aaagaagcag cttcgagaag
     5101 aatcccatcg actttctgga agccaagggc tacaaagaag tgaaaaagga cctgatcatc
     5161 aagctgccta agtactccct gttcgagctg gaaaacggcc ggaagagaat gctggcctct
     5221 gccggcgaac tgcagaaggg aaacgaactg gccctgccct ccaaatatgt gaacttcctg
     5281 tacctggcca gccactatga gaagctgaag ggctcccccg aggataatga gcagaaacag
     5341 ctgtttgtgg aacagcacaa gcactacctg gacgagatca tcgagcagat cagcgagttc
     5401 tccaagagag tgatcctggc cgacgctaat ctggacaaag tgctgtccgc ctacaacaag
     5461 caccgggata agcccatcag agagcaggcc gagaatatca tccacctgtt taccctgacc
     5521 aatctgggag cccctgccgc cttcaagtac tttgacacca ccatcgaccg gaagaggtac
     5581 accagcacca aagaggtgct ggacgccacc ctgatccacc agagcatcac cggcctgtac
     5641 gagacacgga tcgacctgtc tcagctggga ggtgactccg gcggaagctc tggtggcagc
     5701 aagcggaccg ccgacggctc tgaattcgag agccctaaga agaaaagaaa ggtgagcgga
     5761 ggctctagcg gcggaagcgt tatctcctac gacaactacg tgaccatcct ggacgaagag
     5821 acactgaagg cttggatcgc taaactggaa aaagcccctg tgttcgcctt cgccaccgcc
     5881 accgactcct tagataacat cagcgccaac ctggtgggcc tgtctttcgc catcgagcct
     5941 ggcgtggccg cctacatccc cgtggcccac gattacctgg atgccccaga tcaaatcagc
     6001 agagagagag cactggagct cctgaagccc ctgctggagg acgagaaggc cctgaaggtg
     6061 ggacagaacc tgaaatacga taggggaatc ctggctaatt acggcatcga gctgagaggc
     6121 atcgccttcg ataccatgct ggaatcttac atcctgaaca gcgtggccgg ccgccacgac
     6181 atggacagcc tggctgaacg gtggctgaag cacaaaacca tcaccttcga agagatcgcc
     6241 ggaaagggca agaaccagct gacatttaat cagatcgcac tggaagaagc cggcagatac
     6301 gccgctgagg acgccgacgt gacccttcag ctgcacctga agatgtggcc cgatctgcag
     6361 aagcacaagg gcccactaaa cgtgttcgag aacatcgaga tgcctctggt gcccgtcctg
     6421 agcagaatcg agaggaatgg cgtgaagatc gaccccaagg tgctgcacaa ccacagcgag
     6481 gaactgacac tgcggctggc cgagctggaa aagaaggccc acgaaatcgc cggcgaggag
     6541 ttcaacctgt ctagcaccaa gcagctacaa acaatcctgt tcgagaagca gggcattaag
     6601 cctctgaaaa agacacctgg cggagccccc agcaccagcg aagaagtgct ggaggaactg
     6661 gcgctggatt accctctgcc caaagtgatc ctggaatata gaggactggc caagctgaag
     6721 tctacataca ccgacaagct gcctctgatg atcaacccca agaccggccg ggtgcataca
     6781 tcttatcacc aggccgtgac agccaccggc cggctgagca gcacagatcc taacctgcaa
     6841 aacatcccag tgcggaacga agagggcaga agaatcagac aggctttcat tgcccctgag
     6901 gactacgtca tcgtgtctgc tgactacagc cagatagaac tgagaatcat ggcccatctg
     6961 tctcgggaca agggcctgct gaccgctttt gcagaaggca aggacatcca cagagccacc
     7021 gctgccgaag tgttcggcct gcctctggaa acagtgacca gcgagcaaag aaggtccgcc
     7081 aaggccatta acttcggcct gatctacggc atgagcgcct ttggcctggc cagacagctc
     7141 aacatcccta gaaaggaggc ccagaaatac atggacctgt acttcgagcg gtaccccggc
     7201 gtgctggagt acatggaacg gaccagagcc caggctaaag agcagggata cgtggaaacc
     7261 ctggatggcc ggagactgta tctgcctgac atcaagagca gcaacggcgc cagaagagcc
     7321 gccgctgaga gagctgctat caacgcccct atgcagggca ccgccgccga catcatcaag
     7381 cgggccatga tcgccgtgga cgcctggctg caggccgagc agcctagagt gcggatgatc
     7441 atgcaggtgc acgacgagct ggtgtttgag gtgcacaagg acgacgtgga tgctgtggcc
     7501 aagcagattc accagctgat ggaaaattgc accagactgg acgtgccact gctggtcgag
     7561 gttggaagcg gagagaattg ggaccaggcc cactctggcg gctcaaaaag aaccgccgac
     7621 ggcagcgaat tcgagtctcc caagaagaag aggaaagtct gaccggtcat catcaccatc
     7681 accattgagt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg
     7741 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
     7801 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
     7861 gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg
     7921 atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctcg ataccgtcga
     7981 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     8041 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tagggtgcct
     8101 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     8161 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     8221 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     8281 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     8341 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     8401 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     8461 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     8521 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     8581 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     8641 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     8701 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     8761 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     8821 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     8881 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     8941 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     9001 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     9061 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     9121 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     9181 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     9241 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     9301 tgataccgcg cgaaccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     9361 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     9421 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     9481 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     9541 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     9601 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     9661 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     9721 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     9781 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     9841 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     9901 aacccactcg tgcaccca
//
